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Recombinant vegfa

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Recombinant VEGFA is a protein produced using recombinant DNA technology. It is a key regulator of angiogenesis, the process of new blood vessel formation. VEGFA plays a crucial role in various physiological and pathological processes.

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Lab products found in correlation

2 deoxyglucose, by Merck Group (2 mentions) Anti akt, by Cell Signaling Technology (2 mentions) Anti pan actin, by Cell Signaling Technology (2 mentions) Anti fatp3, by Proteintech (2 mentions) Anti fatp4, by Abnova (2 mentions) Sulfo n succinimidyl oleate, by Santa Cruz Biotechnology (2 mentions) Activated charcoal, by Merck Group (2 mentions) Anti calnexin, by Thermo Fisher Scientific (2 mentions) Anti erk1 2, by Cell Signaling Technology (2 mentions) Calcimycin, by Merck Group (2 mentions) 2 4 dinitrophenol, by Merck Group (2 mentions) Anti p erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions) Anti pkc θ, by BD (2 mentions) Anti hibadh, by Proteintech (2 mentions) Anti occludin 1, by Abcam (2 mentions) Vegf b, by R&D Systems (2 mentions) Anti p akt ser473, by Cell Signaling Technology (2 mentions) Mitotracker red cmxros, by Cell Signaling Technology (2 mentions) Matrigel matrix growth factor reduced, by BD (2 mentions) Su6656, by Merck Group (1 mentions)

6 protocols using recombinant vegfa

1

Western Blot and Immunostaining Techniques

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Anti-Pan-Actin (#4968), Anti-ERK1/2 (#9102), anti-pERK1/2 Thr 202/Tyr204 (#9101), anti-AKT (#4685), anti-pAKT Ser473 (#9271 all from Cell Signaling), anti-FATP3 (Proteintech 12943-1-AP), anti-FATP4 (Abnova H00010999-M01), anti-PKC-θ (BD Transduction lab 610089), anti-Na,K ATPase A1 (Novus Biologicals NB300-146SS) and anti-HIBADH (Proteintech 13466-1-AP) antibodies were used for Western blot (1:1000 dilution). Anti-Occludin-1 (Abcam ab31721) and anti-Calnexin (Thermo MA3-27) antibodies were used for immunostaining (1:200 dilution). Mitotracker® Red CMXRos (Cell Signaling 9082S) was used for mitochondria staining. Sulfo-N-succinimidyl Oleate (sc-208408) was purchased from Santa Cruz. Akt VIII (#124018) was purchased from Calbiochem. CHC (#5029) was purchased from Tocris. Recombinant VEGFA (#293-VE-010) and VEGFB (#767-VE-010) were purchased from R&D systems. Human insulin (Humulin R U-100) was purchased from Harvard Drug Group (#821501). Membrane filters (MWCO 3 kDa #Z677094), Calcimycin (#C9275), 2,4-Dinitrophenol (#D198501), 2-deoxyglucose (#D8375), activated charcoal (#C4386) and other chemicals were purchased from Sigma unless otherwise stated.
Jang C., Oh S.F., Wada S., Rowe G.C., Liu L., Chan M.C., Rhee J., Hoshino A., Kim B., Ibrahim A., Baca L.G., Kim E., Ghosh C.C., Parikh S.M., Jiang A., Chu Q., Forman D.E., Lecker S.H., Krishnaiah S., Rabinowitz J.D., Weljie A.M., Baur J.A., Kasper D.L, & Arany Z. (2016). A branched chain amino acid metabolite drives vascular transport of fat and causes insulin resistance. Nature medicine, 22(4), 421-426.
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2

Exploring VEGFR-2 and Integrin Signaling in HUVECs

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Primary human umbilical vein endothelial cells (HUVECs) were purchased from Cascade Biologics. FN, MGO and su6656 were obtained from Sigma. Recombinant VEGF‐A, recombinant RAGE, recombinant human integrin α5β1 protein, RAGE function‐blocking antibody, biotinylated anti‐RAGE antibody, biotinylated anti‐integrin β1 antibody, streptavidin‐HRP conjugate and HRP substrate were from R&D Systems. Growth‐factor‐reduced Matrigel Matrix was from BD Biosciences. Antibodies to phosphorylated VEGFR‐2, total VEGFR‐2, phosphorylated Akt, total Akt, phosphorylated extracellular regulated protein kinases 1/2 (ERK1/2), total ERK1/2, phosphorylated nuclear factor‐κB (NF‐κB), total NF‐κB and CD31 were from Cell Signaling Technology. Antibodies to FN and AGEs were from Abcam. Antibodies to RAGE and c‐Src were from Santa Cruz Biotechnology. Pierce classic IP kit was from Thermo Scientific.
Chen T., Dong J., Zhou H., Deng X., Li R., Chen N., Luo M., Li Y., Wu J, & Wang L. (2020). Glycation of fibronectin inhibits VEGF‐induced angiogenesis by uncoupling VEGF receptor‐2‐c‐Src crosstalk. Journal of Cellular and Molecular Medicine, 24(16), 9154-9164.
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3

Western Blot and Immunostaining Techniques

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Anti-Pan-Actin (#4968), Anti-ERK1/2 (#9102), anti-pERK1/2 Thr 202/Tyr204 (#9101), anti-AKT (#4685), anti-pAKT Ser473 (#9271 all from Cell Signaling), anti-FATP3 (Proteintech 12943-1-AP), anti-FATP4 (Abnova H00010999-M01), anti-PKC-θ (BD Transduction lab 610089), anti-Na,K ATPase A1 (Novus Biologicals NB300-146SS) and anti-HIBADH (Proteintech 13466-1-AP) antibodies were used for Western blot (1:1000 dilution). Anti-Occludin-1 (Abcam ab31721) and anti-Calnexin (Thermo MA3-27) antibodies were used for immunostaining (1:200 dilution). Mitotracker® Red CMXRos (Cell Signaling 9082S) was used for mitochondria staining. Sulfo-N-succinimidyl Oleate (sc-208408) was purchased from Santa Cruz. Akt VIII (#124018) was purchased from Calbiochem. CHC (#5029) was purchased from Tocris. Recombinant VEGFA (#293-VE-010) and VEGFB (#767-VE-010) were purchased from R&D systems. Human insulin (Humulin R U-100) was purchased from Harvard Drug Group (#821501). Membrane filters (MWCO 3 kDa #Z677094), Calcimycin (#C9275), 2,4-Dinitrophenol (#D198501), 2-deoxyglucose (#D8375), activated charcoal (#C4386) and other chemicals were purchased from Sigma unless otherwise stated.
Jang C., Oh S.F., Wada S., Rowe G.C., Liu L., Chan M.C., Rhee J., Hoshino A., Kim B., Ibrahim A., Baca L.G., Kim E., Ghosh C.C., Parikh S.M., Jiang A., Chu Q., Forman D.E., Lecker S.H., Krishnaiah S., Rabinowitz J.D., Weljie A.M., Baur J.A., Kasper D.L, & Arany Z. (2016). A branched chain amino acid metabolite drives vascular transport of fat and causes insulin resistance. Nature medicine, 22(4), 421-426.
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4

Examining Angiogenic Signaling Pathways

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MGO, STZ and human VN were from Sigma (St Louis, MO, USA). Recombinant VEGF-A was from R&D Systems (Minneapolis, MN, USA). Growth-factor-reduced BD Matrigel Matrix (BD Biosciences, San Jose, CA, USA) was from Chemicon International (Temecula, CA, USA). Antibodies to phosphorylated VEGFR-2, total VEGFR-2, phosphorylated Akt, total Akt, phosphorylated ERK1/2, total ERK1/2 and β3 integrin were form Cell Signaling Technology (Beverly, MA, USA). Antibodies to platelet endothelial cell adhesion molecule-1 (PECAM-1) and smooth muscle α-actin were form Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody to AGEs was from Abcam (Cambridge, MA, USA). Antibody to VN was from R&D Systems. LM609 (anti-αvβ3 antibody) and anti-phosphotyrosine antibodies were from Merck Millipore (Watford, UK).
Wang L., Zhang X., Pang N., Xiao L., Li Y., Chen N., Ren M., Deng X, & Wu J. (2015). Glycation of vitronectin inhibits VEGF-induced angiogenesis by uncoupling VEGF receptor-2–αvβ3 integrin cross-talk. Cell Death & Disease, 6(6), e1796-.
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5

HUVEC Migration Assay Using VEGF Chemotaxis

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HUVECs were stained with Cell Tracker Red (Molecular Probes, Eugene, OR, USA) according to the manufacturer’s instruction and seeded in EBM supplemented with 2% FBS in transwell inserts with a FloroBlok membrane with 8-μm pore size in a 24-well plate (Corning, NY, USA). Recombinant VEGF-A (R&D Systems) or conditioned medium containing VEGF-A protein from purified VEGF mRNA (10 ng/mL) in EBM with 2% serum was added to the lower chamber as chemoattractant. After 24 hr, fluorescent microscopy images were obtained.
Carlsson L., Clarke J.C., Yen C., Gregoire F., Albery T., Billger M., Egnell A.C., Gan L.M., Jennbacken K., Johansson E., Linhardt G., Martinsson S., Sadiq M.W., Witman N., Wang Q.D., Chen C.H., Wang Y.P., Lin S., Ticho B., Hsieh P.C., Chien K.R, & Fritsche-Danielson R. (2018). Biocompatible, Purified VEGF-A mRNA Improves Cardiac Function after Intracardiac Injection 1 Week Post-myocardial Infarction in Swine. Molecular Therapy. Methods & Clinical Development, 9, 330-346.
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6

VEGF-A Induced HUVEC Proliferation

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HUVECs were seeded in 96-well plates, at a density of 3,000 cells per well, in EGM (Lonza). The next day, medium was changed to basal EBM supplemented with 2% fetal bovine serum (FBS) with recombinant VEGF-A (R&D Systems) or to conditioned medium containing VEGF-A protein from purified VEGF mRNA. After 48 hr, cells were fixed in 4% buffered formaldehyde (Histolab, Göteborg, Sweden) and nuclei stained with Hoechst (Invitrogen) for 10 min. Fluorescent microscopy images were taken using an automated microscope (ImageXpress Micro XLS; Molecular Devices), and nuclear count was analyzed using MetaXpress software (5.3.0.1; Molecular Devices).
Carlsson L., Clarke J.C., Yen C., Gregoire F., Albery T., Billger M., Egnell A.C., Gan L.M., Jennbacken K., Johansson E., Linhardt G., Martinsson S., Sadiq M.W., Witman N., Wang Q.D., Chen C.H., Wang Y.P., Lin S., Ticho B., Hsieh P.C., Chien K.R, & Fritsche-Danielson R. (2018). Biocompatible, Purified VEGF-A mRNA Improves Cardiac Function after Intracardiac Injection 1 Week Post-myocardial Infarction in Swine. Molecular Therapy. Methods & Clinical Development, 9, 330-346.
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